How to Make Tryptic Soy Agar (TSA)

Step-by-Step Guide

1. 1

   Step 1: Using the marker and tape label the flask with your name

   Continue at high heat and medium high stir rate until all agar is dissolved, approximately 15 minutes. ,,,, (Tip: to determine if the solution is ready pick it up with your hot mitt and hold the flask to the light.
   
   If you see any crystals then the solution is not ready.) ,,, This can be done by placing the rod in the flask and touching the stir-bar. ,, The tape should hold the cap and flask together. ,
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   Step 2: the date and the name of the solution (TSA).

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   Step 3: Carefully slide the large magnetic stir-bar into the Erlenmeyer flask.

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   Step 4: Measure 500 mL of distilled water with the graduated cylinder.

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   Step 5: Pour 500 mL of distilled water from the graduated cylinder into the Erlenmeyer flask.

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   Step 6: Place metal cap on the flask.

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   Step 7: Place flask on the hot stir plate

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   Step 8: turn the plate to high temperature setting and bring to a boil.

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   Step 9: Place the clean plastic weighing boat on the top-loading balance and tare (zero).

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    Step 10: Weigh approximately 6 grams of agar.

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    Step 11: With your hot mitt remove the flask from the hot stir plate and pour agar into the flask.

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    Step 12: Place the metal cap back on the flask and place the flask back on the hot stir plate.

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    Step 13: Turn on the stir plate selection and start mixing at medium speed

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    Step 14: then gradually increase the mixing rate to medium-high over a period of one minute.

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    Step 15: Place the clean plastic weighing boat on the top-loading balance and tare (zero).

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    Step 16: Weigh out approximately 15 grams of TSB.

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    Step 17: With your hot mitt remove the flask from the hot stir plate and pour TSB into the flask.

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    Step 18: Place the metal cap back on the flask and continue to heat the solution on high temperature setting and medium-high stir rate until it is no longer cloudy.

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    Step 19: When the TSA solution is clear

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    Step 20: remove it from the hot stir plate using your hot mitt and place it on a clean surface that doesn’t receive much airflow.

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    Step 21: Turn off the hot stir plate.

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    Step 22: Use the long teflon magnetic tip rod to remove the stir-bar in the flask.

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    Step 23: Cover the flask with the metal cap.

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    Step 24: Cut off a strip of autoclave tape that is approximately 3 inches (7.6 cm) long and place the tape vertically on the side of the metal cap down to the flask.

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    Step 25: Take your solution to the autoclave and follow autoclaving directions.

Detailed Guide

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